Non Animal Testing Database

3D model of the proximal tubule for mechanistic studies

University of Konstanz, Konstanz, Germany
The proximal tubule is the primary site for the reabsorption and secretion of solutes in the kidney, making it a prime target for drug-induced toxicity. 2D cell cultures are unable to fully recapitulate clinical drug responses due to their limited functional lifespan in vitro and the inevitable loss of differentiation when cells are cultured on classical 2D plastic surfaces. Here, the researchers presented the development and characterization of a 3D model of the proximal tubule with primary renal proximal tubule epithelial cells (RPTEC/TERT1 cells). RPTEC/TERT1 cells assembled in Matrigel to form highly differentiated and stable 3D tubular structures characterized by a branched network of single-layered cells enclosing a cell-free lumen, mimicking the proximal tubule. Tubules formed in vitro resembled the polarity of a proximal tubule epithelium, as shown by the polar expression of Na+/K+-ATPase. In addition, 3D-cultured RPTEC/TERT1 cells showed overall increased mRNA expression of xenobiotic transporters, as well as de novo expression of the organic anion transporter OAT3, compared with cultures on plastic or membrane inserts. Finally, this model was used to evaluate delayed cisplatin-induced nephrotoxicity and showed higher sensitivity compared with 2D culture. Thus, the easy-to-use model described here may prove useful for mechanistic studies, such as the discovery of agents affecting tubule formation, differentiation, and polarization, as well as for the detection and understanding of drug-induced nephrotoxicity.
RPTEC/TERT1 cells form highly differentiated tubules when cultured in a 3D matrix
Daniel R. Dietrich
Added on: 11-16-2022
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